Efficient encapsulation of antisense oligonucleotides in lipid vesicles using ionizable aminolipids: formation of novel small multilamellar vesicle structures
Open accessAbstract
Typical methods used for encapsulating antisense oligodeoxynucleotides (ODN) and plasmid DNA in lipid vesicles result in very low encapsulation efficiencies or employ cationic lipids that exhibit unfavorable pharmacokinetic and toxicity characteristics when administered intravenously. In this study, we describe and characterize a novel formulation process that utilizes an ionizable aminolipid (1,2-dioleoyl-3-dimethylammonium propane, DODAP) and an ethanol-containing buffer system for encapsulating large quantities (0.15--0.25 g ODN/g lipid) of polyanionic ODN in lipid vesicles. This process requires the presence of up to 40% ethanol (v/v) and initial formulation at acidic pH values where the DODAP is positively charged. In addition, the presence of a poly(ethylene glycol)-lipid was required during the formulation process to prevent aggregation. The 'stabilized antisense-lipid particles' (SALP) formed are stable on adjustment of the external pH to neutral pH values and the formulation process allows encapsulation efficiencies of up to 70%. ODN encapsulation was confirmed by nuclease protection assays and (31)P NMR measurements. Cryo-electron microscopy indicated that the final particles consisted of a mixed population of unilamellar and small multilamellar vesicles (80--140 nm diameter), the relative proportion of which was dependent on the initial ODN to lipid ratio. Finally, SALP exhibited significantly enhanced circulation lifetimes in mice relative to free antisense ODN, cationic lipid/ODN complexes and SALP prepared with quaternary aminolipids. Given the small particle sizes and improved encapsulation efficiency, ODN to lipid ratios, and circulation times of this formulation compared to others, we believe SALP represent a viable candidate for systemic applications involving nucleic acid therapeutics.
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PEGylated, neutral, and ionizable lipids are employed as stable carriers in lipid delivery systems designed specifically for siRNA.
“PEGylated, neutral, and ionizable lipids are employed as stable carriers in lipid delivery systems designed specifically for siRNA, despite toxicity, complement activation, and poor biological distribution also encouraging their development.317,318”
From Smart nanoparticles for cancer therapy (2023) · cites this paper as [318]
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Self-citation
- Authors
- Semple, S. C., Klimuk, S. K., Harasym, T. O., Dos Santos, N., Ansell, S. M., Wong, K. F., Maurer, N., Stark, H., Cullis, P. R., Hope, M. J., & Scherrer, P.
- Affiliations
- University of British Columbia (CA)
- Philipps University of Marburg (DE)
- Corresponding author
- Semple, Sean C.
- Venue
- Biochimica et Biophysica Acta (BBA) - Biomembranes
- Publisher
- Elsevier BV
- Published
- 2001-02
- Volume
- 1510
- Issue
- 1-2
- Pages
- 152-166
- ISSN
- 0005-2736 (ISSN-L)
- OpenAlex
- W2078775246
- Cited by
- 454
- References
- 55
- Citations by year
- 2026: 15 · 2025: 36 · 2024: 42 · 2023: 38 · 2022: 33 · 2021: 38 · 2020: 18 · 2019: 19 · 2018: 15 · 2017: 8 · 2016: 13 · 2015: 9 · 2014: 21 · 2013: 15 · 2012: 18
- FWCI
- 3.47
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Biochimica et Biophysica Acta (BBA) - Biomembranes (publishedVersion)
- PubMed (publishedVersion)
- Max Planck Institute for Plasma Physics (submittedVersion)
- Max Planck Digital Library (submittedVersion)
- MPG.PuRe (Max Planck Society) (submittedVersion)
- Funders
- Canada Excellence Research Chairs, Government of Canada
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmaceutical Science › Advanced Drug Delivery Systems
- Keywords
- antisense oligonucleotides, lipid vesicles, encapsulation efficiency, particle aggregation, circulation time, nucleic acid therapeutics, lipoplexes
- MeSH terms
- Ethanol, Animals, Cations, Drug Carriers, Female, Hydrogen-Ion Concentration, Liposomes, Membrane Lipids, Mice, Inbred ICR, Magnetic Resonance Spectroscopy, Phosphatidylcholines, Polyethylene Glycols, Single-Strand Specific DNA and RNA Endonucleases, Oligonucleotides, Antisense, Octoxynol, Cryoelectron Microscopy, Mice
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